The UIUC Neuroproteomics and Neurometabolomics Center on Cell-Cell Signaling develops innovative measurement and analysis tools and provides these tools to the NIDA research community. Rapidly evolving metabolomics, peptidomics, and proteomics methods facilitate new findings in both discovery and targeted modes. The Center provides high-end 'omics-scale characterization of the small molecules, peptides, and proteins in samples obtained from brain sub-regions like defined nuclei and even specific single cells. Our sampling methods facilitate molecular localization via high-throughput single cell isolation, mass spectrometry imaging, measurement of activity-dependent release, and quantitation of level changes as a function of exposure to drugs. We can then characterize the most important molecular targets in these samples using metabolomics, peptidomics, and proteomics (bottom-up, middle-down, and top-down) via a broad array of mass spectrometry-based technologies. Finally, we provide the critical expertise for capturing the value of data via expert bioinformatics support that integrates disparate data types, develops advanced analytical approaches for complex metabolomics and proteomics experiments, and provides community support through several web platforms. At the beginning of the next project period, we will be supporting an initial group of 19 major users representing the fields of addiction research and neuroscience, including projects targeting neuropeptides, transmitters, proteins, and protein complexes that are involved in multiple aspects of drug escalation, exposure and addiction, and the interaction of neuropeptides, pain pathways, and opioids. We also will address fundamental questions of neuron/astrocyte identity, the first comprehensive single cell lipid profiles of defined cell types, and even measure drug exposure as a function of time using hair samples (to aid several ongoing NIDA trials). The Neuroproteomics and Neurometabolomics Center on Cell-Cell Signaling is divided into three research cores: the Sampling and Separation Core, the Molecular Profiling and Characterization Core, and the Bioinformatics, Data Analytics and Predictive Modeling Core (plus an Administrative Core). New this year, we have added a Pilot Research Project Core that invites new users to interact with our Center, and have outlined an exemplary group of three initial pilot projects that include a beginning investigator and a new direction for an established PI. The high level of synergy between the neuroscientists and technologists affiliated with the Center ensures we will enable exciting scientific advances in understanding how systems of neurons interact in both the healthy nervous system and upon exposure to drugs of abuse. Lastly, a series of outreach initiatives confirms that our protocols and approaches are widely available to the appropriate scientific communities.